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为明确miRNA沉默靶基因的调控机理,克隆AGO1蛋白基因并了解其作为沉默复合物核心成员在miRNA沉默通路中的作用,本研究在分析‘金冠’苹果基因组的基础上,以‘金冠’叶片cDNA为模板,克隆获得了3 234bp AGO1基因全长,蛋白分子质量为119ku,等电点为9.41,包含2个保守的AGO蛋白特征结构域:PAZ和Piwi区,命名为MdAGO1。苹果‘金冠’不同组织MdAGO1基因及13种与发育相关miRNA实时定量PCR分析结果表明:1)MdAGO1在花和果实中表达量均高于叶片;2)miRNA在果实和花中表达量均较高,叶片相对较低。MdAGO1与13种发育相关miRNA表达一致,在苹果中MdAGO1可能与miRNA协同作用调控植物的生长发育。
In order to clarify the regulatory mechanism of miRNA silencing target genes, AGO1 gene was cloned and its function as a core member of silencing complex in miRNA silencing pathway was analyzed. Based on the analysis of ’Golden Delicious’ apple genome, As a template, the 3 234 bp AGO1 gene was cloned. The molecular mass of the gene was 119 ku and the isoelectric point was 9.41. It contained two conserved AGO protein domains: PAZ and Piwi, named as MdAGO1. The results of real-time quantitative PCR analysis of MdAGO1 gene and 13 miRNAs related to development showed that: 1) The expression of MdAGO1 in flowers and fruits was higher than that in leaves; 2) The expression of miRNA was higher in fruits and flowers , The leaves are relatively low. MdAGO1 is consistent with 13 developmental miRNAs. In apple, MdAGO1 may cooperate with miRNA to regulate plant growth and development.